B型利鈉肽對(duì)心肌細(xì)胞線粒體動(dòng)力學(xué)的作用研究
發(fā)布時(shí)間：2019-06-27 12:01
【摘要】： 心肌缺血再灌注(ischemia/reperfusion, I/R)損傷是缺血心肌在恢復(fù)血流后可引起心肌超微結(jié)構(gòu)、功能、代謝等各方面的進(jìn)一步損害,并存在著不同程度心肌細(xì)胞凋亡的現(xiàn)象,是臨床十分常見(jiàn)的病理過(guò)程。導(dǎo)致心肌缺血再灌注損傷的機(jī)制是多方面的,氧化應(yīng)激、細(xì)胞內(nèi)Ca2+超載、線粒體損傷和中性粒細(xì)胞活化都參與了心肌結(jié)構(gòu)與功能的改變。如何減輕再灌注造成的損傷,已成為當(dāng)前心血管疾病研究領(lǐng)域的熱點(diǎn)問(wèn)題。而線粒體的結(jié)構(gòu)和功能改變?cè)谛募∪毖俟嘧p傷中起關(guān)鍵作用。線粒體是真核細(xì)胞中重要的細(xì)胞器,因其在能量代謝和細(xì)胞凋亡的關(guān)鍵作用,線粒體形態(tài)結(jié)構(gòu)和功能動(dòng)力學(xué)研究已成為心血管疾病研究熱點(diǎn)。最新研究發(fā)現(xiàn)線粒體內(nèi)外膜上分布著一些通道,這些結(jié)構(gòu)在細(xì)胞損傷時(shí)可能被激活并發(fā)揮一定生物學(xué)效應(yīng),甚至可以決定細(xì)胞的生死存亡。這些通道主要有線粒體通透性轉(zhuǎn)換孔(mitochondrial permeability transition, MPTP)線粒體鈣轉(zhuǎn)運(yùn)體(mitochandrial Ca2+ uniporter, MCU)和線粒體ATP敏感鉀離子通道(mitochandrial ATP-sensitive potassium channel,MitoKATP)等。此外線粒體是高度動(dòng)態(tài)的細(xì)胞器,線粒體的結(jié)構(gòu)和形態(tài)動(dòng)力學(xué)都將決定線粒體功能,甚至參與到細(xì)胞凋亡的信號(hào)通路中。有研究提示缺血再灌損傷時(shí)伴有線粒體結(jié)構(gòu)和形態(tài)動(dòng)力學(xué)的改變,而其中的作用機(jī)制還需進(jìn)一步探討和研究。 B型利鈉肽(B-type natriuretic peptide, BNP)是利鈉肽家族中一種重要的生物活性物質(zhì),具有強(qiáng)大的排鈉、利尿、舒張血管和抑制腎素—醛固酮系統(tǒng)激活等作用,已是心力衰竭等心臟疾病的重要生物標(biāo)記物。BNP可作用于心肌細(xì)胞上分布的相應(yīng)鈉肽受體,可激活鳥(niǎo)苷酸環(huán)化酶產(chǎn)生環(huán)磷酸鳥(niǎo)苷參與細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)通路,產(chǎn)生相應(yīng)的生物學(xué)效應(yīng)。人工重組BNP已經(jīng)美國(guó)SFDA批準(zhǔn)可用于治療急性失代償性心力衰竭。最新研究提示BNP可通過(guò)作用于線粒體途徑來(lái)減輕心肌缺血再灌注損傷,發(fā)揮保護(hù)心肌細(xì)胞結(jié)構(gòu)和功能作用,并且BNP這種保護(hù)作用可能與線粒體ATP敏感鉀離子通道有關(guān)。BNP在臨床上治療心力衰竭和其他心臟疾病的研究和評(píng)價(jià)還不明確,尤其BNP對(duì)心肌缺血再灌損傷的作用機(jī)制和作用靶點(diǎn)研究國(guó)內(nèi)外鮮有報(bào)道。 為了闡明BNP參與心肌細(xì)胞中和線粒體相互的作用機(jī)制,本研究以離體培養(yǎng)的心肌細(xì)胞缺血再灌模型和心肌H9c2細(xì)胞系細(xì)胞骨架破壞模型作為實(shí)驗(yàn)對(duì)象,從細(xì)胞和分子兩個(gè)水平研究BNP對(duì)線粒體的功能動(dòng)力學(xué)和形態(tài)運(yùn)動(dòng)動(dòng)力學(xué)作用機(jī)制,并對(duì)之相關(guān)的信號(hào)通路進(jìn)行了探討。本研究包括以下內(nèi)容： 首先建立離體培養(yǎng)的心肌細(xì)胞缺血再灌模型,采用了流式細(xì)胞術(shù)、激光共聚焦顯微鏡和蛋白免疫印跡等手段,實(shí)驗(yàn)結(jié)果提示BNP可減輕缺血再灌損傷導(dǎo)致的心肌細(xì)胞凋亡,抑制線粒體膜電位的丟失和活性氧的生成,并可以抑制Bax和Smac/DIABLO凋亡蛋白的表達(dá),減少線粒體cytochrome c的釋放,增加Bcl-2抗凋亡蛋白的表達(dá)。結(jié)果提示BNP在心肌缺血再灌過(guò)程中可能通過(guò)抑制線粒體通透性轉(zhuǎn)換孔和線粒體鈣轉(zhuǎn)運(yùn)體的開(kāi)放,從而發(fā)揮保護(hù)心肌細(xì)胞作用,而B(niǎo)NP的這些作用與通過(guò)激活PI3K-Akt信號(hào)通路有關(guān)。 其次建立了心肌細(xì)胞H9c2細(xì)胞骨架破壞模型,通過(guò)細(xì)胞活性檢測(cè),細(xì)胞指數(shù)實(shí)時(shí)測(cè)定和細(xì)胞成像等研究手段和方法,證明了BNP可以保護(hù)心肌細(xì)胞的線粒體功能和形態(tài)運(yùn)動(dòng)動(dòng)力學(xué),可減輕細(xì)胞骨架微管破壞導(dǎo)致的心肌細(xì)胞凋亡,抑制線粒體膜電位的丟失和活性氧的生成,并可以抑制線粒體cytochrome c的生成和釋放,維持線粒體的空間分布和運(yùn)動(dòng)。 綜上所述,本研究的主要發(fā)現(xiàn)和結(jié)論是：1、BNP在心肌缺血再灌過(guò)程中可通過(guò)抑制線粒體通透性轉(zhuǎn)換孔和線粒體鈣轉(zhuǎn)運(yùn)體的開(kāi)放,維持線粒體功能動(dòng)力學(xué),減輕心肌細(xì)胞損傷；2、BNP可激活PI3K-Akt信號(hào)通路,抑制細(xì)胞內(nèi)線粒體相關(guān)凋亡通路,保護(hù)心肌細(xì)胞；3、BNP可通過(guò)穩(wěn)定細(xì)胞骨架微管成分來(lái)維持線粒體形態(tài)結(jié)構(gòu)動(dòng)力,保護(hù)心肌細(xì)胞。本研究為BNP用于臨床心臟疾病的治療和闡明線粒體與心臟疾病關(guān)系提供了實(shí)驗(yàn)研究依據(jù)。
[Abstract]:The myocardial ischemia-reperfusion (I/ R) injury is the further damage of the myocardial ultrastructure, function, metabolism and other aspects of the ischemic myocardium after the blood flow is restored, and the phenomenon of the apoptosis of the cardiac muscle cells in different degrees is a very common pathological process. The mechanism of myocardial ischemia-reperfusion injury is multiple, oxidative stress, intracellular Ca2 + overload, mitochondrial injury and neutrophil activation are involved in the changes of myocardial structure and function. How to reduce the damage caused by reperfusion has become a hot topic in the research field of cardiovascular disease. The structural and functional changes of mitochondria play a key role in myocardial ischemia-reperfusion injury. Mitochondria is an important organelle in eukaryotic cells, because of its key role in energy metabolism and cell apoptosis, the study of mitochondrial morphological structure and functional dynamics has become a hot topic in the study of cardiovascular disease. The most recent study found that there are some channels on the inner and outer membrane of the mitochondria, which can be activated and exert a certain biological effect when the cells are damaged, and can even determine the survival and death of the cells. These channels mainly have the mitochondrial permeability transition (MPTP) mitochondrial calcium transporter (MPTP), the mitochondrial ATP-sensitive potassium channel (MCU) and the mitochondrial ATP-sensitive potassium channel (Mitogen ATP). In addition, the mitochondria are highly dynamic organelles, and the structure and morphology of the mitochondria will determine the mitochondrial function and even participate in the signal pathway of cell apoptosis. There are changes in the structure and morphology of the mitochondria in the case of ischemia-reperfusion injury, and the mechanism of the mechanism is further discussed and studied. The B-type natriuretic peptide (BNP) is an important bioactive substance in the natriuretic peptide family. It is an important biological marker of heart disease such as heart failure and the like, which has the effects of sodium excretion, diuresis, vasodilation and the inhibition of the activation of the renin-aldosterone system. The BNP can act on the corresponding sodium peptide receptor distributed on the cardiac muscle cell, and can activate the ornithine acid cyclase to generate the ring-type ornithine to participate in the cell signal transduction pathway to produce the corresponding biology. Effect. Artificial recombinant BNP has been approved by the FDA for the treatment of acute decompensation The latest study suggests that BNP can reduce the myocardial ischemia-reperfusion injury by acting on the mitochondrial pathway, play a role in protecting the structure and function of the myocardial cells, and the protective effect of BNP may be related to the mitochondrial ATP-sensitive potassium ion channel. The research and evaluation of BNP in the treatment of heart failure and other cardiac diseases is not clear, especially the mechanism and target of BNP for myocardial ischemia-reperfusion injury. In order to clarify the mechanism of the interaction of BNP in the myocardial cells and the mitochondria, the model of the myocardial ischemia-reperfusion model and the cell skeleton destruction model of the myocardial H9c2 cell line were studied in this paper. In order to study the dynamic and dynamic mechanism of BNP on the function and morphology of the mitochondria from two levels of cell and molecule, the relevant signal pathway was studied. A discussion is made. This research package The experimental results suggested that BNP can reduce the damage of ischemia-reperfusion injury. The apoptosis of myocardial cells is inhibited, the loss of mitochondrial membrane potential and the generation of active oxygen are inhibited, the expression of Bax and Smac/ DIABLO apoptosis proteins can be inhibited, the release of the mitochondrial cytochrome c is reduced, and the Bcl-2 is increased. The results suggest that BNP may play a role in protecting the cardiac muscle cells by inhibiting the opening of the mitochondrial permeability transition hole and the mitochondrial calcium transporter during the myocardial ischemia/ reperfusion, and these effects of the BNP are related to the activation of PI3K-A. It is proved that BNP can protect the line of cardiac muscle cells by means of cell activity detection, cell index real-time measurement and cell imaging. the kinetic of body function and morphology can reduce the apoptosis of the myocardial cells caused by the destruction of the cytoskeleton microtubule, inhibit the loss of mitochondrial membrane potential and the generation of active oxygen, and can inhibit the generation and release of the mitochondrial cytochrome c, To sum up, the main findings and conclusions of this study are:1. The main findings and conclusions of this study are:1. BNP can inhibit the open of the mitochondrial permeability transition hole and the mitochondrial calcium transporter in the course of myocardial ischemia and reperfusion, maintain the mitochondrial function dynamics and reduce the myocardial cell damage;2, the BNP can The PI3K-Akt signaling pathway is activated to inhibit the mitochondria-related apoptosis pathway in the cell and to protect the cardiac muscle cells; and 3, the BNP can maintain the line particles by stabilizing the cytoskeleton microtubule component. The present study is the treatment and elucidation of the mitochondria and the heart for the treatment of clinical heart diseases.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R341
本文編號(hào)：2506770
[Abstract]:The myocardial ischemia-reperfusion (I/ R) injury is the further damage of the myocardial ultrastructure, function, metabolism and other aspects of the ischemic myocardium after the blood flow is restored, and the phenomenon of the apoptosis of the cardiac muscle cells in different degrees is a very common pathological process. The mechanism of myocardial ischemia-reperfusion injury is multiple, oxidative stress, intracellular Ca2 + overload, mitochondrial injury and neutrophil activation are involved in the changes of myocardial structure and function. How to reduce the damage caused by reperfusion has become a hot topic in the research field of cardiovascular disease. The structural and functional changes of mitochondria play a key role in myocardial ischemia-reperfusion injury. Mitochondria is an important organelle in eukaryotic cells, because of its key role in energy metabolism and cell apoptosis, the study of mitochondrial morphological structure and functional dynamics has become a hot topic in the study of cardiovascular disease. The most recent study found that there are some channels on the inner and outer membrane of the mitochondria, which can be activated and exert a certain biological effect when the cells are damaged, and can even determine the survival and death of the cells. These channels mainly have the mitochondrial permeability transition (MPTP) mitochondrial calcium transporter (MPTP), the mitochondrial ATP-sensitive potassium channel (MCU) and the mitochondrial ATP-sensitive potassium channel (Mitogen ATP). In addition, the mitochondria are highly dynamic organelles, and the structure and morphology of the mitochondria will determine the mitochondrial function and even participate in the signal pathway of cell apoptosis. There are changes in the structure and morphology of the mitochondria in the case of ischemia-reperfusion injury, and the mechanism of the mechanism is further discussed and studied. The B-type natriuretic peptide (BNP) is an important bioactive substance in the natriuretic peptide family. It is an important biological marker of heart disease such as heart failure and the like, which has the effects of sodium excretion, diuresis, vasodilation and the inhibition of the activation of the renin-aldosterone system. The BNP can act on the corresponding sodium peptide receptor distributed on the cardiac muscle cell, and can activate the ornithine acid cyclase to generate the ring-type ornithine to participate in the cell signal transduction pathway to produce the corresponding biology. Effect. Artificial recombinant BNP has been approved by the FDA for the treatment of acute decompensation The latest study suggests that BNP can reduce the myocardial ischemia-reperfusion injury by acting on the mitochondrial pathway, play a role in protecting the structure and function of the myocardial cells, and the protective effect of BNP may be related to the mitochondrial ATP-sensitive potassium ion channel. The research and evaluation of BNP in the treatment of heart failure and other cardiac diseases is not clear, especially the mechanism and target of BNP for myocardial ischemia-reperfusion injury. In order to clarify the mechanism of the interaction of BNP in the myocardial cells and the mitochondria, the model of the myocardial ischemia-reperfusion model and the cell skeleton destruction model of the myocardial H9c2 cell line were studied in this paper. In order to study the dynamic and dynamic mechanism of BNP on the function and morphology of the mitochondria from two levels of cell and molecule, the relevant signal pathway was studied. A discussion is made. This research package The experimental results suggested that BNP can reduce the damage of ischemia-reperfusion injury. The apoptosis of myocardial cells is inhibited, the loss of mitochondrial membrane potential and the generation of active oxygen are inhibited, the expression of Bax and Smac/ DIABLO apoptosis proteins can be inhibited, the release of the mitochondrial cytochrome c is reduced, and the Bcl-2 is increased. The results suggest that BNP may play a role in protecting the cardiac muscle cells by inhibiting the opening of the mitochondrial permeability transition hole and the mitochondrial calcium transporter during the myocardial ischemia/ reperfusion, and these effects of the BNP are related to the activation of PI3K-A. It is proved that BNP can protect the line of cardiac muscle cells by means of cell activity detection, cell index real-time measurement and cell imaging. the kinetic of body function and morphology can reduce the apoptosis of the myocardial cells caused by the destruction of the cytoskeleton microtubule, inhibit the loss of mitochondrial membrane potential and the generation of active oxygen, and can inhibit the generation and release of the mitochondrial cytochrome c, To sum up, the main findings and conclusions of this study are:1. The main findings and conclusions of this study are:1. BNP can inhibit the open of the mitochondrial permeability transition hole and the mitochondrial calcium transporter in the course of myocardial ischemia and reperfusion, maintain the mitochondrial function dynamics and reduce the myocardial cell damage;2, the BNP can The PI3K-Akt signaling pathway is activated to inhibit the mitochondria-related apoptosis pathway in the cell and to protect the cardiac muscle cells; and 3, the BNP can maintain the line particles by stabilizing the cytoskeleton microtubule component. The present study is the treatment and elucidation of the mitochondria and the heart for the treatment of clinical heart diseases.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R341
【引證文獻(xiàn)】
相關(guān)博士學(xué)位論文 前1條
1 盧娜;葒草素對(duì)缺血再灌注損傷心肌細(xì)胞的保護(hù)作用及相關(guān)機(jī)制研究[D];浙江大學(xué);2011年
，本文編號(hào)：2506770
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